A Service for Human Chromosome Studies in Saskatchewan

A service has been developed in Saskatchewan to make available the results of studies of human chromosomes, the material being forwarded to the laboratory by local transport facilities. During the first year of this project chromosome studies were requested for five doubtful cases of trisomy-21 (two were found to be normal) and for 20 definite cases of trisomy-21 in young patients (two had translocations but the parents of both these children had normal karyotypes). Eleven confirmed cases of Turner''s syndrome, two of Klinefelter''s syndrome, and one each of the D and E syndromes were also studied. The largest group for which studies were requested comprised 36 patients with mental retardation; only two abnormal karyotypes were encountered in this group.     

水稻、小麦、油菜和烟草细胞质雄性不育系统中组分Ⅰ蛋白的比较分析

组分Ⅰ蛋白(RuBP羧化酶/加氧酶)的生物合成系由叶绿体基因和细胞核基因共同控制,所以,被作为研究细胞质遗传的标记。本实验用免疫化学和氨基酸成分分析等方法,对水稻(珍汕97)、小麦(繁7)、油菜(湘矮早)和烟草(G28)的细胞质雄性不育系及其保持系的组分Ⅰ蛋白作了比较,同时对不同作物的组分Ⅰ蛋白也作了免疫鉴定。结果表明,细胞质雄性不育系及其保持系的组分Ⅰ蛋白差异不大,但是,四种不同作物的组分Ⅰ蛋白之间有明显差异。     

Nucleotide sequence and expression of cheF, an essential gene for chemotaxis in Bacillus subtilis.

The cheF gene, which is involved in chemotaxis in Bacillus subtilis, has been cloned, expressed, and sequenced. This gene is contained in a 0.7-kilobase PstI DNA fragment that was isolated from a lambda Charon 4A B. subtilis chromosomal DNA library. This fragment was subcloned into the expression vector pSI-1 and shown to complement the cheF mutation both for chemotaxis and for methanol production in response to the addition of attractants. Plasmid-encoded DNA expression in B. subtilis maxicells indicated that a membrane-associated polypeptide of 20-kilodaltons was expressed from this 0.7-kilobase DNA. The nucleotide sequence of this DNA fragment was determined, and an open reading frame capable of encoding a putative 175-amino-acid protein (Mr 20,002) was identified. In an effort to understand the function of the cheF protein, the dosage of the cheF gene product was varied by altering the concentration of IPTG (isopropyl-beta-D-thiogalactopyranoside) during growth. In the presence of high concentrations of IPTG, chemotaxis was inhibited and methanol production was impaired.     

Tyrosine-7 is an essential residue for the catalytic activity of human class PI glutathione S-transferase: chemical modification and site-directed mutagenesis studies.

The glutathione (GSH)-conjugating activity of human class Pi glutathione S-transferase (GST pi) toward 1-chloro-2,4-dinitrobenzene (CDNB) was significantly lowered by reaction with N-acetylimidazole, an O-acetylating reagent for tyrosine residues. Further, the replacement of Tyr7 in GST pi, which is conserved in all cytosolic GSTs, with phenylalanine by site-directed mutagenesis also lowered the activities toward CDNB and ethacrynic acid. The Km values of the mutant for both GSH and CDNB were almost equivalent to those of the wild type, while the Vmax of the former was about 55-fold smaller than that of the latter. Therefore, Tyr7 is considered to be an essential residue for the catalytic activity of GST pi.     

不同底物与固氮酶及其钼铁蛋白相互作用的荧光光谱分析

 本文研究了不同底物(N_2,H_2,N_2O,NaN_3,C_2H_2)对棕色固氮菌固氮酶及其钼铁蛋白荧光光谱的影响。结果表明,上述底物均能络合在钼铁蛋白及固氮酶上,但络合程度不同,从而为固氮酶系统有多个不同的底物络合中心,底物络合中心在钼铁蛋白分子上,铁蛋白对钼铁蛋白有变构作用,提供了光谱学证据。     

槐种子发育中胚乳细胞半乳甘露聚糖积累的研究

槐 ( Sophora japonica L.)开花约 60 d至种子成熟 ,为胚乳半乳甘露聚糖积累期。用组织化学方法 ,对储藏于胚乳细胞壁上的半乳甘露聚糖的形成积累进行了观察 ,结果表明 ,半乳甘露聚糖最先在邻近胚的胚乳细胞的粗面内质网的囊泡腔内形成 ,并通过细胞质膜分泌至细胞壁周围。此后 ,半乳甘露聚糖的积累逐渐向种皮方向扩展 ,及至种子成熟时 ,除糊粉层外 ,所有胚乳细胞几乎全由多糖所填充。此外 ,对半乳甘露聚糖发生部位及其积累过程的消长变化进行了讨论     

Distinct tyrosine phosphorylation sites in ZAP-70 mediate activation and negative regulation of antigen receptor function.

Biochemical and genetic evidence has implicated two families of protein tyrosine kinases (PTKs), the Src- and Syk-PTKs, in T- and B-cell antigen receptor signaling. ZAP-70 is a member of the Syk-PTKs that associates with the T-cell antigen receptor and undergoes tyrosine phosphorylation following receptor activation. Three tyrosine residues, Tyr-292, -492, and -493, have been identified as sites of phosphorylation following T-cell antigen receptor engagement. Utilizing ZAP-70- and Syk-deficient lymphocytes (Syk-DT40 cells), we provide biochemical and functional evidence that heterologous trans-phosphorylation of Tyr-493 by a Src-PTK is required for antigen receptor-mediated activation of both the calcium and ras pathways. In contrast, cells expressing mutations at Tyr-292 or -492 demonstrate hyperactive T- and B-cell antigen receptor phenotypes. Thus, phosphorylation of ZAP-70 mediates both activation and inactivation of antigen receptor signaling.     

Single-stranded DNA binding protein and DNA helicase of bacteriophage T7 mediate homologous DNA strand exchange.

Two proteins encoded by bacteriophage T7, the gene 2.5 single-stranded DNA binding protein and the gene 4 helicase, mediate homologous DNA strand exchange. Gene 2.5 protein stimulates homologous base pairing of two DNA molecules containing complementary single-stranded regions. The formation of a joint molecule consisting of circular, single-stranded M13 DNA, annealed to homologous linear, duplex DNA having 3'- or 5'-single-stranded termini of approximately 100 nucleotides requires stoichiometric amounts of gene 2.5 protein. In the presence of gene 4 helicase, strand transfer proceeds at a rate of > 120 nucleotides/s in a polar 5' to 3' direction with respect to the invading strand, resulting in the production of circular duplex M13 DNA. Strand transfer is coupled to the hydrolysis of a nucleoside 5'-triphosphate. The reaction is dependent on specific interactions between gene 2.5 protein and gene 4 protein.     

Theory of multivalent binding in one and two-dimensional lattices

Ligand binding to a linear lattice composed of N sites, under general conditions of cooperativity and number of sites covered upon binding, m, is approached in terms of the theory of contracted partition functions. The partition function of the system obeys a recursion relation leading to a generating function that provides an exact analytical solution for any case of interest. Site-specific properties of the lattice are derived from simple transformations of the analytical expressions. The McGhee-von Hippel model is obtained as a special case in the limit N --> infinity. The derivation is straightforward and involves no combinatorial arguments. Partition functions and site-specific properties are also derived for the case of non-cooperative binding to a two-dimensional torus of length N, containing s sites in its section for a total of sN sites. The torus provides a relevant model for ligand binding to double-stranded DNA (s = 2) or protein helices (s = 3,4). It is proved that non-cooperative binding to the two-dimensional torus can mimic cooperative binding to a one-dimensional linear lattice when m = s. The dimensional embedding of the lattice and the geometry of interaction of its sites play a crucial role in defining the binding properties of the system accessible to experimental measurements. Hence, caution must be exercised in the interpretation of Scatchard plots in terms of the one-dimensional McGhee-von Hippel model, especially when m < or = 4 and the geometry of the system is clearly two-dimensional.     

丝孢酵母高甲硫氨酸突变株的选育及营养调控

以丝孢酵母(Trichosporon Behr)ST851为原始菌株,经紫外线诱变,在含乙硫氨酸的双层平板上筛选到多株抗乙硫氨酸突变株。其中ST851-10株抗乙硫氨酸浓度达到350μg/ml,其菌体蛋白质含量由40.5％提高到44.3％,菌体甲硫氨酸含量由20.45mg/g-DCW增加到29.32mg/g-DCW。在以苹果渣为碳源、尿素为氮源、硫酸镁作硫源的最适培养条件下,固态发酵24h后,蛋白质和甲硫氨酸含量较原始菌株分别提高了15.8％和44.9％。培养基中C/N值低有利于甲硫氨酸的合成,C/N值高则适合于菌体生长。在苹果渣固态发酵过程中,适当补加氮源既有利于菌体生长和甲硫氨酸的合成,又可起到调节培养基pH值的作用。